International Journal of ChemTech esearch CDE (USA): IJCGG ISS: 0974-4290 Vol.8, o.4, pp 2096-2100, 2015 Synthesis and Antimicrobial Activity of 2-amino-4- (substitutedphenyl)-6-{4-[3-chloro-2-(4-hydroxyphenyl)-4- Shailesh.H.Shah 1 * 1 Department of Chemistry, Patel JB Arts, Patel AM Commerce & Patel JDKD Science College, Borsad-388540, Gujarat, India Abstract: Pyridine the simplest and perhaps is the best-known heterocyclic compound. The credit for the discovery of pyridine goes to Anderson who first obtained it from bone oil. The simple pyridine compounds are prepared by the Cyclization of aliphatic raw materials. Preparation of 3-cyanopyridines is available in the literature with different methods1-5. A new series of 2-amino-4- (substitutedphenyl)-6-{4-[3-chloro-2-(4-hydroxyphenyl)-4-oxoazetidin -1-yl] phenyl} pyridine-3-carbonitrile are synthesized by reacting 3-chloro-1-{4-[5-(Substituted phenyl)-4,5- dihydro-pyrazol-3-yl]phenyl}-4-(4-hydroxyphenyl)azetidin-2-one with malanonitrile and ammonium acetate by using ethanol as a solvent. All these compounds were characterized by means of their I, 1 H M, Spectroscopic data and microanalysis. All the compounds were tested for their antibacterial and antifungal activities by broth dilution method. Keywords: Chalcones, Cyanopyridine, azetidin-2-one, Antimicrobial activity. Introduction: Interest in the synthesis of pyridine containing compounds has increased in recent years because of their biological and pharmacological activities. Pyridine is the parent compound of the series of compounds that is important in pharmaceutical, agriculture and industrial chemistry. Among a wide range of pyridines 3- cyanopyridines acquired a special attention due to their wide range of therapeutic activities. Some substituted pyridine-3-carbonitrile and their derivatives have been reported to possess some interesting biological activities such as Antifungal 6, Antiepileptic 7, Antibacterial 8, Anticonvulsant 9, Anti tubercular 10, Analgesic 11, Insecticidal 12, Antipsoriasis 13, Antihypertensive 14.. In the present study we report the reaction of 3-chloro-1-{4-[5-(Substituted phenyl)-4, 5-dihydropyrazol-3-yl] phenyl}-4-(4-hydroxyphenyl) azetidin-2-one with malanonitrile and ammonium acetate to form pyridine-3-carbonitrile (4a-j). The structures of the various synthesized compounds were assigned on the basis of I, 1 H-M spectral data and elemental analysis. These compounds were also screened for their antimicrobial activity. Experimental: The I spectra were recorded on I affinity-1, DS-8000A, Shimadzu, Ptc. Ltd., Japan spectrophotometer. The 1 H-M was recorded in DMS on Bruker Advance II 400 MHz spectrometer using
Shailesh.H.Shah /Int.J. ChemTech es. 2015,8(4),pp 2096-2100. 2097 TMS as an internal standard. Melting points were determined in open capillary tubes and are uncorrected. The purity of the compounds was checked by TLC-using Silica gel-g (Merck). Column chromatography was performed on silica gel. All the compounds were tested for their antibacterial and antifungal activities by broth dilution method eaction Scheme H H 2 methanol H Et 3 CCH 2 1 H H 3a-j H 2 H 2 Ethanol 2 Preparation of 1-(4-{[(4-hydroxyphenyl) methylene] amino} phenyl) ethanone (1) A mixture of 4-hydroxy benzaldehyde (0.01M), 1-(4-aminophenyl) ethanone (0.01M) and methanol (30ml) was heated for about 5 min. in a beaker (250 ml) to get a clear solution. The solution was kept overnight at room temperature to get the respective crude solid which was recrystallized from ethanol to obtain the pure crystals of 1-(4-{[(4-hydroxy phenyl)methylene]amino}phenyl)ethanone respectively. The yield of the product was 75% and the product melts at 195 0 C. : C(75.28%) H(5.45%) (5.82%), Calcd. for C 15 H 13 2 : C(75.30%) H(5.48%) (5.85%). I,cm -1 :3085 (-H), 3040 (=C-H), 2920(-C-H), 1676(>C=), 1647(>C=-), 1606 (>C=C<), 1363(-, bend), 1314(-C-<), 1284 (-C--), 1240(-C-C-C-). 1 H-M (DMS, δ, ppm): 2.5692 (3H, s, C ), 6.5277-7.9774 (8H, m, Ar-H), 8.3820 (1H, s, -CH=-), 9.6392 ( 1H, s, Ar-H). Preparation of 1-(4-acetylphenyl)-3-chloro-4-(4-hydroxyphenyl) azetidin -2-one (2) In a 100ml ound bottom flask 1-(4-{[(4-hydroxyphenyl) methylene] amino} phenyl) ethanone (0.01M) in 70ml benzene was taken. Chloro acetyl chloride (0.01M) was added at room temperature with constant stirring and triethylamine 1ml was added and the reaction mixture was refluxed for 7 hours. After the completion of reaction, solvent was removed by vacuum distillation. The solid was filtered, dried and recrystallized from toluene. The yield of the product was 60% and the product melts at 119 0 C. : C(64.64%) H(4.44%) (4.42%), Calcd. for C 17 H 14 3 : C(64.67%) H(4.47%) (4.44%). I, cm -1 : 3300 (- H), 3050(=C-H), 2950(-C-H), 1680(>C=), 1600(>C=C<), 1375(-, bend), 1300(-C-<), 1240(-C-C-C- ), 1220(-C-), 560 (C-). 1 H-M (DMS, δ, ppm): 2.5392 (3H, s, C ), 4.8954 (1H, d, >CH-Ar), 5.5151 (1H, d, >CH-), 6.6720-8.0745 (8H, m, Ar-H), 9.7784 ( 1H, s, Ar-H). Preparation of 3-chloro-1-{4-[3-(Substituted phenyl) prop-2-enoyl] phenyl}-4-(4-hydroxyphenyl) azetidin -2-one (3a-j) To the solution of 1-(4-acetylphenyl)-3-chloro-4-(4-hydroxyphenyl) azetidin-2-one (0.01M) in absolute ethanol (50 ml), substituted benzaldehyde (0.01M) and 2% ah were added and refluxed for 10 hours. After refluxing the reaction mixture was concentrated, cooled, filtered and neutralized with dil. H. The solid residue thus obtained was crystallized by absolute ethanol. I(3b), cm -1 :3359(-H), 3045(=C-H), 1728(>C=), 1608(>C=C<), 1290(-C-<), 1186 (-C--), 769(-C-). 1 H-M (3c-DMS, δ, ppm): 3.8789 (6H, s, - ), 4.8613 (1H, d, >CH-Ar), 5.3413 (1H, d, >CH-), 6.7340-7.8883 (11H, m, Ar-H), 7.9733 (2H, d, -CH=CH-), 9.8306 ( 1H, s, Ar-H).
Shailesh.H.Shah /Int.J. ChemTech es. 2015,8(4),pp 2096-2100. 2098 eaction Scheme H 4 C H 2 C H 3a-j CH 2 C 2 H 4a-j Preparation 2-amino-4-(substitutedphenyl)-6-{4-[3-chloro-2-(4-Hydroxyphenyl)-4-oxoazetidin-1-yl] phenyl} pyridine-3-carbonitrile.(4a-j) A mixture of 3-chloro-1-{4-[3-(Substituted phenyl) prop-2-enoyl] phenyl}-4-(4-hydroxyphenyl) azetidin-2-one (0.01M) in 30 ml alcohol then 0.011 Mole of malanonitrile and 0.06 Mole of ammonium acetate was added and refluxed for 7 h. Then the resulting product was cooled into crushed ice, filtered, washed by H2, dried and recrystallized by ethanol. I(4j), cm -1 : 3358 (-H), 3040 (=C-H), 1693(>C=), 1658(>C=-), 1581 (>C=C<), 1372(-, bend), 1324(-C-<),1284 (--), 1215 (-C-), 1075 (-C--C-). Table: 1 Physical constant of 2-amino-4-(substitutedphenyl)-6-{4-[3-chloro-2-(4-Hydroxyphenyl)-4- Compd M.F. 4a Yie ld % M.P. C % C C 27 H 18 2 4 2 68 215 64.63 (64.68) Elemental Analysis % 11.14 (11.17) % H 3.59 (3.62) 4b H C 27 H 19 4 3 65 180 67.11 (67.15) 11.57 (11.60) 3.92 (3.97) 4c C 29H 23 4 4 75 206 66.07 (66.10) 10.58 (10.63) 4.37 (4.40) 4d 2 C 27 H 18 5 4 60 230 63.31 (63.35) 13.64 (13.68) 3.50 (3.54) 4e C 27 H 18 2 4 2 70 210 64.62 (64.68) 11.14 (11.17) 3.58 (3.62) 4f 4g CH 2 C 31 H 28 5 2 65 217 69.17 (69.20) CH 2 H C 27 H 19 4 3 73 168 67.11 (67.15) 12.98 (13.02) 11.55 (11.60) 5.21 (5.25) 3.91 (3.97) 4h 4i C 29 H 24 5 2 63 205 68.27 (68.30) C 27 H 19 4 2 69 202 69.41 (69.45) 13.69 (13.73) 11.95 (12.00) 4.70 (4.74) 4.06 (4.10) 4j H C 28 H 21 4 4 64 215 85.51 (65.56) 10.88 (10.92) 4.09 (4.13)
Shailesh.H.Shah /Int.J. ChemTech es. 2015,8(4),pp 2096-2100. 2099 esults and discussion Antimicrobial activity The MICs of synthesized compounds were carried out by broth micro dilution method as described by atan (2000). It is one of the non automated in vitro bacterial susceptibility tests. This classic method yields a quantitative result for the amount of antimicrobial agents that is needed to inhibit growth of specific microorganisms. The invitro antimicrobial activity of test compounds were assessed against 24 hr cultures of several selected bacteria and fungi. The bacteria used were E. coli, S.aureus, P. aeruginosa, and S. pyogenus; the fungi used were C. albicans, A. iger, and A.clavatus. The antimicrobial activity was performed by broth dilution method in DMS. Gentamycin, Ampicilin, Chloramphenicol, Ciprofloxacin, orfloxacin, ystatin and Greseofulvin were used as standard for the evaluation of antibacterial and antifungal activities respectively. The activity was reported by Minimal Inhibition Concentration. The results are summarized in Table-2 Table: 2 Antimicrobial activity of 2-amino-4-(substitutedphenyl)-6-{4-[3-chloro-2-(4-Hydroxyphenyl)-4- S.. CMP.. ATIBACTEIAL ACTIVITY MIIMAL IHIBITI CCETATI ATIFUGAL ACTIVITY MIIMAL IHIBITI CCETATI E.CLI P.AEUGISA S.AUEUS S.PYGEUS C.ALBICAS A.IGE A.CLAVATUS MTCC 443 MTCC 1688 MTCC 96 MTCC 442 MTCC 227 MTCC 282 MTCC 1323 1 4a -2-500 500 500 500 500 500 >1000 2 4b -2-H 250 100 100 175 1000 500 500 3 4c -3-, -4-250 500 100 500 1000 250 250 4 4d -3-2 125 100 100 62.5 500 500 500 5 4e -4-250 125 100 175 1000 >1000 500 6 4f -4-(C 2 H 5 ) 2 200 100 62.5 100 1000 500 500 7 4g -4-H 500 250 125 200 500 500 250 8 4h -4-( ) 2 100 100 100 125 1000 500 500 9 4i -H 250 100 500 100 >1000 500 1000 10 4j -3-, -2-H 100 500 250 500 500 500 1000 Table: 3 Antibacterial Activity: Minimal Inhibition Concentration (The Standard Drugs) Drug E.Coli P.Aeruginosa S.Aureus S.Pyogenus - MTCC 443 MTCC 1688 MTCC 96 MTCC 442 (MICGAMME/ML) Gentamycin 0.05 1 0.25 0.5 Ampicillin 100 -- 250 100 Chloramphenicol 50 50 50 50 Ciprofloxacin 25 25 50 50 orfloxacin 10 10 10 10 Table: 4 Antifungal Activity: Minimal Inhibition Concentration (The Standard Drugs) Drug C.Albicans A.iger A.avatus - MTCC 227 MTCC 282 MTCC 1323 (MICGAMME/ML) ystatin 100 100 100 Greseofulvin 500 100 100
Shailesh.H.Shah /Int.J. ChemTech es. 2015,8(4),pp 2096-2100. 2100 Biological screening result of activities 2-amino-4-(substitutedphenyl)-6-{4-[3-chloro-2-(4-Hydroxy phenyl)-4- based derivatives shows that compound (4d, 4h & 4j) have shown better activity against E. coli and (4f & 4i ) against S. pyogenus, while (4j) have Shawn better activity against S. Aureus, while rest of all compound possessed good activity against S.aureus in the range of 62.5-500 µg/ml. Compound ( 4a, 4d, 4g & 4j) is found to be good antifungal activity against C. albicans, against standard drugs Greseofulvin. While rest of all derivatives are poor against A. iger, and A.clavatus Conclusion The Main focus of this research work was to synthesize, characterize and evaluate antimicrobial activities of the newly synthesized pyridine-3-carbonitrile derivatives, structures of synthesized compounds were confirmed and characterized with the help of analytical data s such as I and 1 H-M. In summary, we have described the synthesis and antimicrobial activity of some new 2-amino-4-(substitutedphenyl)-6-{4-[3- chloro-2-(4-hydroxyphenyl)-4- MIC values revealed that amongst newly synthesized compound having itro and 3-methoxy,2- hydroxy type linkage has shown good activity against the bacterial strains.. Acknowledgement The author are thankful to the Principal and Management of Patel JB Arts, Patel AM Commerce & Patel JDKD Science College, Borsad for providing laboratory facilities, SAIF, Chandigarh for M Spectra and Loyola esearch Center- Xavier s College, Ahmedabad for I spectra and micro-care laboratory, Surat, Gujarat, India for biological activity. eferences 1. Crossley M. L., King V. L., orthey L. H. and Scholz T. E., (1949) US 2, 491, 253; (1961) Chem. Abstr., 45, 4746 2. Krivokolysko S. G., (1999) Chem. Heterocylc. Comp (. Y.) 3. Dayochenko U. P., (1998) uss. J. rg. Chem., 34(4), 554-556; (1999) Chem.Abstr. 130, 223222c 4. Sayed G. H., Kassab.., (1998) Bull. Fac. Pharm., (1999) Chem. Abstr. 131, 15727p 5. kazoe Takashi, (2002) PCT Int. Appl. W 0006, 547; Chem. Abstr., 132, 321784y 6. Latif., Mishrky. and Girgis. S., (1981) Indian J. Chem., 20B, 147-149 7. Von Behenburg W., Engel J., Heese J. and Thiele K., (1984) Ger. ften.,d.e., 3, 337, 593 ( C07D 213/72) ; (1984) Chem. Abstr., 101, 130595n 8. Castedo L., Quintela J. M. and iguers., (1984) Eur. J. Med. Chem., 19(6), 555; (1985) Chem. Abstr., 103, 37337 9. Pavia M.., Taylor C. P., Hershenson F. M. and Lobbestael S. J., (1987) J. Med Chem., 30, 1210 10. Hoefling W. L., Elhaner D. and eckling E., (1965) VEB Leund-Werke Walter Ulbricht Ger. 1, 193, 506; (1965) Chem. Abstr., 63, 6979 11. Thiele Kurt, Von Be Benburg and Walter E., (1970) S. African. J. Chem., 6, 905-906 12. John B., Freeman and Peter F. M., (1971) Ger. ften. 2, 029, 079 (. A 01 007d); (1969) Brist. Appl., (1971) Chem. Abstr. 74, 99891d 13. Scott V. and Joseph, (1979) Jap. Pat., 2, 803, 592; (1980) Chem. Abstr, 92, 47216 14. Baldwin J. J., Scrialrine A., Ponticeello G. S., Engelhardt E. L. and Sweeti C. S.,(1980) J. Heterocycl. Chem., 17(3), 425; (1980) Chem. Abstr, 93, 186222 *****